【作者】 于济洋；

【导师】 李新华；

【作者基本信息】 沈阳农业大学 ， 食品科学与工程， 2014， 博士

【摘要】 菊芋(Jerusalem artichoke),俗称鬼子姜、洋姜。鲜菊芋中含有丰富的营养物质,包括碳水化合物为16.6%(其中78%为菊糖,又称菊芋多糖),此外,还含有少量蛋白质、粗纤维、氨基酸以及多种矿物质。菊芋种植分布广,耐寒旱耐贫瘠,在我国有广阔的栽种面积,在辽宁省内的阜新、大连瓦房店和鞍山岫岩地区更具有广泛的种植基础,是-种非常具有开发潜力的宝贵资源。在菊糖的加工过程中需要脱蛋白、除杂等复杂工艺,制备成本高,而且获得的产品中可能会有化学试剂的残留,导致菊糖使用时可能会有一定的安全隐患。将不易贮存的鲜菊芋经脱皮、干燥、粉碎后制成以菊糖为主要成分的菊芋全粉,即避免了加工溶剂的残留,保留了菊芋中大量的营养物质,又能更好地解决鲜菊芋存储和运输的问题。因此,与菊糖相比较,菊芋全粉的研究开发更具有现实意义。研究发现,菊糖具有非常好的加工特性,在食品工业中可以作为增稠剂替代品、脂肪替代品及保湿剂,广泛应用于速冻肉制品、发酵乳制品和糕点等食品的加工过程中,不仅能改变食品的流变及质构特性,而且还具有膳食纤维和促进益生菌增殖等生理功能。以往的研究主要集中在菊糖上,而对于菊芋全粉的研究则未见报道,因此,菊芋全粉能否替代菊糖的某些重要加工特性,需要进行比较分析后才能得出结论。本文通过比较分析菊芋全粉与菊糖的基本性质,进一步了解菊芋全粉的加工特性,同时分别对两种重要加工特性凝胶特性和发酵特性进行了基础研究,在菊芋全粉特性研究基础上,对其进行改性,利用改性后菊芋全粉具有的螯合特性,制备出螯合钙产品,并对其增加钙吸收的效果和机理进行研究,既为菊芋全粉的应用提供了技术支持,又实现了菊芋产品高附加值的合理利用,研究项目具有一定的社会效益、经济效益,更有利于提高人民健康水平,对推进我国高效农业和绿色食品的发展具有重要意义。主要结论如下：一、在辽宁大连地区种植的三种主要菊芋品种中,辽东小红更适合用作生产菊糖的原料。通过热风干燥并粉碎制得的菊芋全粉中,淀粉、蛋白质、脂肪、灰分和粗纤维含量比菊糖中相应含量高,纯菊糖含量为73.70±3.65%,比菊糖中纯菊糖含量低14%左右,但菊芋全粉中纯菊糖仍占主要成分,自制的菊芋全粉呈淡黄色、粉末状。菊芋全粉和菊糖虽然二者的基本成分相差较大,但凝胶指数、凝胶时间、保水性、透光性、冻融稳定性、粘度在一定程度上差异不大,二者在一定程度上可以相互替代。菊芋全粉的持油性要好于菊糖,差异极显著(P<0.001),可以应用到含油脂原料的加工食品中。二、菊芋全粉具有良好的凝胶性。其主要凝胶参数即凝胶强度、黏性、弹性、黏聚性、咀嚼性和恢复性随菊芋全粉浓度增加而增大,增加菊糖浓度是改善菊芋全粉体系凝胶性能的有效手段。凝胶强度和黏性随菊芋全粉浓度的提高呈线性关系,菊芋全粉浓度≤50%时,凝胶强度变化较显著；菊芋全粉浓度≤60%时,咀嚼性变化较显著。其余参数受菊芋全粉浓度影响差异不显著。放置时间对凝胶质构性能影响不显著,放置温度可以显著影响全粉的凝胶性能,当温度介于60℃～80℃范围时,全粉形成凝胶。酸、糖、盐能显著影响菊芋全粉凝胶的形成,当pH=6时,凝胶强度达到最大,比自然条件下全粉凝胶体系(pH=6.6)时凝胶强度提高了8%；多糖与二糖和单糖相比,改善全粉凝胶性能的作用更明显；低浓度的食盐可以促进全粉的凝胶性,而高浓度食盐促进凝胶性的增幅有所降低。三、菊芋全粉分别对乳酸菌、双歧杆菌、保加利亚乳杆菌和嗜酸乳杆菌具有良好的发酵性,其做为乳酸菌培养基的增值效果是菊糖的2倍,是葡萄糖的9.7倍；作为碳源对双歧杆菌的增殖效果是菊糖的5.3倍,是葡萄糖的16.8倍；作为碳源对保加利亚乳杆菌的增殖效果是菊糖的4.8倍,是葡萄糖的9.6倍；作为碳源对嗜酸乳杆菌的增殖效果是菊糖的3.7倍,是葡萄糖的9.3倍；菊糖对嗜热链球菌的发酵性较差,使用菊糖作为培养基碳源对嗜热链球菌起不到增殖的效果,但是在菊芋全粉作为碳源的培养基中,嗜热链球菌生长状况与MC培养基的生长状况相差不大。通过单因素试验和响应面曲面分析确定乳酸菌在菊芋全粉替代葡萄糖培养基的最佳发酵条件为：碳源的浓度为2.1%,发酵温度为38℃,接种量为10%,实际测得发酵后的乳酸菌的菌落总数为2.48×109cfu/mL。四、制备羧甲基菊芋全粉的因素中,碱化时间、氢氧化钠与菊芋全粉质量比、一氯乙酸与菊芋全粉质量体积比对羧甲基取代度的影响效果显著,最佳工艺条件为：一氯乙酸与菊芋全粉质量比为1.6：1,氢氧化钠与菊芋全粉质量比为1.1：1,醚化时间3h,此时取代度为1.3。制备羧甲基菊芋全粉螯合钙过程中,溶液pH值、CaCl2与羧甲基菊芋全粉的质量比和螯合时间三个因素对取代度有显著影响,最佳螯合工艺为pH值7,CaCl2与羧甲基菊芋全粉的质量比9：100,螯合时间30min,此时螯合率为63.7%。不同取代度的羧甲基菊芋全粉螯合钙离子的能力不同,在低取代度范围内(0.2<DS <0.67),钙离子螯合率随着取代度的增加迅速提高,在中取代度范围内(0.67<DS<1.05),螯合率继续增加,但增幅不大,在高取代度范围内(1.05<DS<1.27),螯合率呈现下降趋势。五、液相质谱和取代度测试可知,羧甲基菊芋全粉螯合钙可能有三种螯合形式,即果糖内螯合、菊芋全粉分子内螯合和菊芋全粉分子间螯合,果糖内螫合结构所占比例很低,螯合以后两种结构为主。红外光谱测试可知,羧甲基特征主体峰1596cm-1和1425.14cm-1,分别分裂成1608.34cm-1和1562.06cm-1及1450.21cm-1和1425.14cm-1两个特征吸收峰；核磁质谱测试羧甲基化及螯合钙并未改变菊糖的主体结构,羧甲基菊芋全粉螯合钙仍为呋喃型多糖,C-3、C-4和C-6上的羟基具有反应活性,均能被羧甲基取代,羰基碳化学位移由螯合前的175.1变为螯合后的181.9；X-射线衍射测试可知,螯合钙离子后,2θ=5.3°、10.3°和10.7°等处出现新的尖锐衍射峰,7.6°、25.4°和27.2°处衍射峰仍存在,而8.2°、21.3°和29.7°处衍射峰强度较羧甲基菊芋全粉而言减弱；电子显微镜测试可知,羧甲基菊芋全粉螯合钙离子后,丧失了原来的形貌,颗粒表面出现了很多白色钙晶粒“镶嵌”在羧甲基菊糖表面；差式量热扫描和热重分析可知,羧甲基菊芋全粉螯合钙后,热力学性质发生了变化,钙离子螯合量为4.99mmol/g (20.01mg/g)。六、采用双侧卵巢摘除的方法,给成年雌性大鼠去势,建立大鼠骨质疏松模型并以骨质疏松模型大鼠作为载体进行研究,模型组大鼠双侧卵巢摘除10周后,股骨重量、血钙含量、尿钙UCa含量、股骨骨密度FTD指标全部低于空白对照组,且差异显著,组织切片发生减少。从模型组大鼠股骨组织病理切片的形态学改变和测定结果可判断,该模型复制成功。菊芋全粉钙组股骨干重、湿重、血钙、骨密度均显著高于高钙片组,尿钙含量显著低于高钙片组。对大鼠股骨进行切片,切片中较模型组骨小梁明显增多、变粗,骨小梁表面破骨细胞也较少,而成骨细胞较模型组显著增多、增生活跃。羧甲基菊芋全粉螯合钙可有效提高骨质疏松模型大鼠血钙水平,增高骨密度,减轻骨质疏松,其对钙制剂的吸收度优于传统碳酸钙制剂。七、采用羧甲基菊芋全粉螯合钙干预肠道菌群失调模型小鼠,建立肠道菌群失调小鼠模型。通过测试可知,羧甲基菊芋全粉螯合钙具有调节肠道菌群的功能,可以改善小鼠的活动状态,使活动度较低小鼠的精神状态转好,使水样便转为正常；可以使模型组小鼠血清中SOD活性显著上升,MDA活性显著下降；可以降低模型小鼠肠道大肠杆菌DNA水平,升高模型小鼠肠道乳酸菌DNA水平。对于在大肠内钙才被吸收的羧甲基菊芋全粉螯合钙,提高机体抗氧化能力和调整大肠内菌群平衡是可能促进钙离子吸收的机制之一。更多还原

【Abstract】 Jerusalem artichoke, known as Helianthus tuberosus, is rich in nutrients, including16.6%carbohydrate (among which78%are inulin) when it is fresh. In addition, it contains a small amount of protein, crude fiber, amino acids, vitamin and a variety of minerals. Inulin is widely-grown, highly-adaptable, frigostable, drought-enduring and barren-tolerable. It has been broadly planted in China. It is especially suitable for being planting in Fuxin, Wangfangdian of Dalian and Xiulian of Anshan. It’s a precious resource with huge development potential.The processing of inulin needs such complicated processes as deproteinization and edulcoration, which makes its production cost pretty high. In addition, the product obtained has residual chemical reagent, which makes the use of inulin a certain kind of security risk. After decrustation, desiccation and smash, the difficult-to-be-preserved fresh inulin will become inulin granules, the essential component of which is inulin, which both avoids residual solvent from process, maintains lots of nutrients of inulin and better resolve the storage and transportation problem of inulin. So, compared with inulin, the research and development of inulin granules has more practical significance.According to research, inulin is extremely processable. It can be used as a replacement of thickener, fat and humectants and is broadly used in processing of frozen mean product, fermented dairy products and pastry etc. It can not only change rheological and texture characteristics of food but also has such physiological functions as promoting proliferation of probiotics and that dietary fiber has. Previous study mainly focused on inulin and we have not yet seen any reports of studies about inulin granules. So, it should be compared and analyzed in order to get the conclusion of whether inulin granules can be used to replace some major processing characteristics of inulin. The processing characteristics of inulin granules are further studied and the gel and fermentative property are fundamentally studied in this paper through the comparison of basic characteristics between inulin granules and inulin. Based on studied characters of inulin granules, its characters have been changed. The chelating character of inulin granules, after character change, is used to make chelated calcium product. Calcium absorption increase effect and mechanism of calcium preparation which use inulin granules as its chelate substrate is also studied, which not only provide technical support for the application of processing character of inulin granules but also reasonably applied high added value of jerusalem artichoke products. The research project has certain social benefit, economy benefit and benefits the improvement of people’s health level. It is of great importance in promoting the development of highly-effective agriculture and green products. The major conclusions are as follows:I. Among the three major kinds of jerusalem artichokes which are planted in Dalian of Lioaning province, Liaodong Xiaohong is more suitable for being used as the raw material of inulin. Hot air dried and smashed inulin granules has higher content of starch, protein, fat, ash and crude fiber. Synanthrin content in inulin granules is73.70±3.65, nearly14%lower than that in inulin. However, synanthrin is still the major content of inulin granules. Self-made inulin granules is faint yellow and in powder form. Also basic content of inulin granules differs from that of inulin largely, such characters as VGI, gelation time, water holding capability, transmittancy, freeze-thaw stability and viscosity are similar, which makes the two to be used to replace each other. Oil holding capability of inulin granules is better than that of inulin, or we can say their difference is substantial (P<0.001). Inulin granules can be used in the processing of fatty ray materials.II. Inulin granules have better gel capacity. Its major gel capacity parameters, like gel strength, adhesiveness, elasticity, cohesiveness, chewiness and estorative increases with density of inulin granules. Increasing density of inulin granules is an effective way to increase gel capacity of inulin granules system. Gel capacity depends linearly on mass fraction of inulin granules. When inulin granules’density is equal or lesser than50%, gel capacity changes substantially. When inulin granules’density is equal or lesser than60%, chewiness changes substantially. Other parameters are not evidently influenced by inulin granules’ density. Texture performance of inulin granules is not evidently influenced by storage time. Storage temperature can affect gel capacity of inulin granules. When the temperature ranges from60℃to80℃, inulin granules gels. Acid, salt and sugar can affect the formation of inulin granules gel substantially. When PH=6, the intensity of gel reaches its peak, an improvement of6%when compared with the intensity of naturally-formed (pH=6.6) gel of inulin granules system. Polysaccharide and disaccharide, compared with monosaccharide. affects gel capacity of inulin granules more. Low concentration salt affects gel capacity of inulin granules largely while high concentration salt’s impact on gel capacity is small.III. Inulin granules have higher fermentation performance on lactic acid bacteria, bifidobacterium, lactobacillus bulgaricus and lactobacillus acidophilus. Its propagation effect is2fold of that of Inulin and9.7fold of that of glucose when used as lactic acid bacteria substrate. Its propagation effect on bifidobacterium is5.3fold of that of inulin and16.8fold of that of glucose when used as carbon source. Its propagation effect on lactobacillus bulgaricus is4.8fold of that of inulin and9.6fold of that of glucose when used as carbon source. Its propagation effect on lactobacillus acidophilus is3.7fold of that of inulin and9.3fold of that of glucose when used as carbon source. Inulin has poor fermentation performance on streptococcus thermophilus. Inulin has no propagation effect on streptococcus thermophilus when used as substrate carbon source. Growth status of streptococcus thermophilus in substrate carbon source of inulin granules has small difference than that in MRS substrate. Through single factor experiment and respond surface analysis, fermentation conditions of lactobacillus, when glucose substrate is replaced with inulin granules substrate, are determined:concentration of carbon source is2.1%, fermentation temperature is38℃, inoculation size is10%and actual measured fermented total lactobacillus count is2.48×109cfu/mL.Ⅳ. Among all the factors of the processing of CMIG, basification time, quantity ratio between sodium hydroxide and inulin granules, quantity and size ratio between MAC and inulin granules have substantial effect on degree of carboxymethyl substitution. The optimal processing condition is:quantity ratio between monochloroacetic acid and inulin granules is1.6:1; quantity ratio between sodium hydroxide and inulin granules is1.1:1; etherification time is3hours, based on which the substitution degree is1.3. During the process of CMIG production, the following three factors:solution pH value, quantity ratio between CaCl2and CMIG, chelation time, have significant influence on substitution degree. The optimal chelation process is:pH value is7, quantity ratio between CaCl2and carboxymethyl inulin granules is9:100, chelation time is30minutes, based on which the chelation degree is63.7%. Chelating calcium ion capacity of CMIG with different substitution degree differs from each other:in lower substitution degree range (0.2<DS<0.67), chelation rate of calcium ion increases sharply with substitution degree; in middle substitution degree range (0.67<DS <1.05), chelation rate keeps increasing but by a lower increase rate while in higher substitution degree range (1.05<DS<1.27), chelation rate tends to decrease.V. CMIGC may have three chelation forms:chelation within fructose, chelation within inulin granules and chelation between inulin granules. According to liquid mass spectrometry test, percentage of chelation within fructose structure is low and most chelation belongs to the later two structures. According to infrared spectroscopy test,1596cm-1and1425.14cm-1characteristic peaks of carboxymethyl divide into two characteristic absorption peaks:1608.34cm-1and1562.06cm-1as well as1450.21cm-1and1425.14cm-1. According to NMR and MS test, carboxymethylation and chelated calcium have not changed major structure of synanthrin. CMIGC is still furanoid polysaccharose. Oxhydryl in C-3, C-4and C-6is reactive and can be replaced by carboxymethyl. Chemical shift of carbonyl carbon changes from175.1(before chelation) to181.9(after chelation). According to x-ray diffraction test, after chelating calcium ion, sharp diffraction peak will show when2θis5.3°,10.3°and10.7°etc. Diffraction peaks still exists when20is7.6°,25.4°and27.2°. Diffraction peak intensity decreases when8.2°,21.3°and29.7°, compared with CMIG According to electron microscope test, morphological structure of CMIG disappears after it chelates calcium ion. Granules surface see tremendous of calcium grains which inlay in the surface of CMI. According to Differential Scanning Calorimeter and Thermogravimetric Analysis, thermodynamic property of CMIG changes after it chelates calcium ion. Chelation amount of calcium ion is4.99mmol/g(20.01mg/g).Ⅵ. Castrating adult female rat through bilateral ovariectomy method to build rat osteoporosis model and use osteoporosis model rat as carrier. After the application of bilateral ovariectomy on rats in model group, femur quality, blood calcium content, UCa content and femur intesntiy FTD indicator are all lower than that of rats in blank control group and the difference is significant. Tissue slice sees pathologic changes as osteocyte decrease. According to morphological changes and measure results of femur tissue pathological slice of rats in model group, the copy of this model is a success. Dry weight, wet weight, blood calcium and bone mineral density of femur in inulin granules calcium group are higher than that in high calcium group while urinary calcium content is lower than that in high calcium group. Slice femur of rats and bone trabecula in the slice increases significantly and thickens, compared with that in model group, and has less osteoclast on the surface of bone trabecula, while osteoblast is higher in volume and more active in proliferation than that in model group. CMIGC can effectively improve blood calcium level, bone mineral density and relieve osteoporosis of osteoporosis model rat. Its absorbance of calcium preparation is superior to traditional carbonic acid calcium preparation.Ⅶ. Use CMIGC to intervene rats in intestinal bacilli illness group to establish intestinal bacilli illness rats model. According to test, CMIGC can adjust intestinal flora, improve active state of rat and make those inactive rats in good spirit, turn watery stool into normal; enhance SOD activity of rat’s serum and lower MDA content; lower DNA level of colibacillus inside intestinal tract of rats in model group. In terms of CMIGC which can only be absorbed inside large intestine, improving antioxidation ability and adjusting flora balance inside large intestine is one of the mechanisms that is likely to help calcium ion absorption.更多还原